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This highlights the need for delivery carriers capable of concurrently achieving cytosolic delivery and autophagy inhibition. Here, we report a stepwise-coordinated polymeric nanoplatform based on a tailor-made amphiphilic triblock copolymer (mPEG-b-PGBA-b-PGCQ) that enables traceless cytosolic delivery of functional proteins while concurrently suppressing autophagy. This system incorporates three modular blocks: an mPEG stealth shell, a PGBA block for reversible protein conjugation via pH-sensitive iminoboronates, and a PGCQ block grafted with autophagy inhibitor hydroxychloroquine (HCQ) via self-immolative disulfide linkages. Using ribonuclease A (RNase A) as a model therapeutic, we demonstrated efficient protein release in acidic endolysosomes, simultaneous HCQ-promoted endosomal escape, and glutathione-triggered HCQ liberation in cytosol. The released HCQ significantly enhanced RNase A-induced apoptosis by inhibiting both the autophagic degradation of the protein and RNase A-induced mitophagy. Furthermore, by integrating cytosolic delivery with autophagy inhibition, this platform produced notable functional enhancement across multiple proteins, including green fluorescent protein, \u03b2-galactosidase, and horseradish peroxidase, demonstrating its broad utility. Our work establishes a potent traceless delivery and autophagy-blockade strategy for boosting intracellular protein efficacy, with considerable potential in both basic research and biomedical applications.<\/jats:p>","DOI":"10.1093\/nsr\/nwag446","type":"journal-article","created":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T11:39:11Z","timestamp":1784374751000},"source":"Crossref","is-referenced-by-count":0,"title":["Stepwise-coordinated polymeric nanoplatform enables traceless cytosolic protein delivery and autophagy blockade for enhanced tumor therapy"],"prefix":"10.1093","volume":"13","author":[{"given":"Peng","family":"Zhang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Polymer Science and Technology, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun 130022 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaowei","family":"Zhu","sequence":"additional","affiliation":[{"name":"College of Chemistry and Chemical Engineering, Taishan University , Tai\u2019an 271000 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongyu","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Polymer Science and Technology, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun 130022 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuai","family":"Tang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University , Changchun 130012 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhihui","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Polymer Science and Technology, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun 130022 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